Title: Nanoapertures for AFM-based single-molecule force spectroscopy

Authors: Stephan F. Heucke; Elias M. Puchner; Stefan W. Stahl; Alexander W. Holleitner; Hermann E. Gaub; Philip Tinnefeld

Addresses: Center for Nanoscience and Department of Physics, Ludwig-Maximilians-Universität München, Amalienstr. 54, 80799 Munich, Germany ' Department of Cellular and Molecular Pharmacology, University of California, 600 - 16th Street, San Francisco, CA 94158, USA ' Center for Nanoscience and Department of Physics, Ludwig-Maximilians-Universität München, Amalienstr. 54, 80799 Munich, Germany ' Walter Schottky Institut and Physik-Department, Technische Universität München, Am Coulombwall 4a, 85748 Garching, Germany ' Center for Nanoscience and Department of Physics, Ludwig-Maximilians-Universität München, Amalienstr. 54, 80799 Munich, Germany ' Physikalische und Theoretische Chemie - NanoBioSciences, Technische Universität Braunschweig, Hans-Sommer-Strasse 10, 38106 Braunschweig, Germany

Abstract: Simultaneous single-molecule force spectroscopy and microfluorescence binding measurements are often hampered by background fluorescence from the bulk. Zero-Mode Waveguides (ZMW) restrict the excited volume but require a special design, which allows the tip of the force probing cantilever to protrude into the nanoaperture. Here, we describe the design and fabrication of such ZMW and report the first experiments where binding of fluorescent adenosine triphosphate to the force-activated enzyme titin kinase was measured while the enzyme was subjected to mechanical forces.

Keywords: single-molecule force spectroscopy; microfluorescence; zero-mode waveguide; ZMW; nanoapertures; force-activated enzyme; mechanoenzymatics; titin kinase; AFM; atomic force microscopy; nanotechnology; single-molecule fluorescence; fluorescent adenosine triphosphate.

DOI: 10.1504/IJNT.2013.053529

International Journal of Nanotechnology, 2013 Vol.10 No.5/6/7, pp.607 - 619

Received: 08 May 2021
Accepted: 12 May 2021

Published online: 26 Apr 2013 *

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